[No public or meaningful name is available]

Endpoint summary

Description of key information

Skin:

in vivo skin irritation according to OECD 404: slightly irritating, full reversibility

in vitro skin corrosion according to OECD 431: non-corrosive

 

Eye:

in vivo eye irritation according to OECD 405: slightly irritating, full reversibility

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

2007-10-22 to 2007-10-29

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)

Version / remarks:

13 April 2004

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.40 (In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test (TER))

Version / remarks:

April 2005

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Justification for test system used:

EpiDerm™ human skin model can be used for distinguishing between corrosive (C) and non-corrosive (NC) chemicals, giving an excellent prediction for a wide spectrum of chemieals, shows reproducible results and provides good in vivo - in vitro correlation.

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm™ (MatTek)
- Tissue batch number: Lot 8859 Kit c
- Date of initiation of testing: 2007-10-29

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during exposure: 37 ± 1 °C

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: the tissue was gently rinsed about 20 times with PBS (phospate buffered saline) to remove any residual test item.
- Observable damage in the tissue due to washing: no


MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER EXPOSURE
- MTT concentration: not specified
- Incubation time: 3 h
- Spectrophotometer: yes
- Wavelength: 550 nm

NUMBER OF REPLICATE TISSUES:

NUMBER OF INDEPENDENT EXPERIMENTS TO DERIVE FINAL PREDICTION: 1

PREDICTION MODEL
Corrosivity potential of the test item was predicted from the relative mean tissue viabilities obtained after 3 min treatment compared to the negative control tissues concurrently treated with A. dest (= 100%). A test item is classified "corrosive" (C) in any case, if the relative tissue viability after 3 min treatment is decreased below 50%. In addition, those test items classified as "non-corrosive" (NC) after 3 min (viability >= 50%) are classified as "corrosive" if the relative mean tissue viability after 60 min treatment compared to the concurrent negative control tissues is decreased below 15%.

Mean tissue viability Prediction C / NC
(% negative control)

3 min: < 50% C

3 min: >= 50% C
and 60 min: < 15%

3 min: >= 50% NC
and 60 min: >= 15%


ACCEPTANCE CRITERIA
The test meets acceptance criteria if:
- mean OD550 nm of the two negative control tissues of the 3 min and 60 min treatment period is >= 0.8,
- mean relative tissue viability of the two positive contral tissues of the 3 min treatment period is <= 30%,
- maximum inter tissue viability difference between two tissues treated identically is <= 30%.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount applied: 50 µL

VEHICLE
- Amount applied: 50 µL

NEGATIVE CONTROL
- Amount applied: 50 µL

Duration of treatment / exposure:

3 and 60 min

Duration of post-treatment incubation (if applicable):

3 h

Number of replicates:

2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

60 min

Value:

104

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3 min

Value:

107

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

The mixture of 100 µL test item per 1 mL MTT medium showed no reduction of MTT compared to the solvent. The OD550 was 0.3934 (solvent control: 0.6851).

The test item showed no corrosive effects. The mean relative tissue viability (% negative contral) was >= 50% (107%) after 3 min treatment and >= 15% (104%) after 60 min treatment.
The contrals confirmed the validity of the study. The mean OD550 of the two negative control tissues was >= 0.8 for each exposure period. The mean relative tissue viability (% negative control) of the positive contraI was <= 30% (12%) after 3 min treatment. The maximum inter tissue difference of replicate tissues of all dose groups was <= 30%.

Interpretation of results:

GHS criteria not met

Conclusions:

In this study under the given conditions the test item showed no corrosive effects. The relative mean tissue viability after 3 min treatment was >= 50% and after 60 min. treatment >= 15%. The test item is therefore classified as "non corrosive".

Executive summary:

The skin corrosivity potential of the test item was analysed according to OECD 431. Since corrosive chemicals are cytotoxic after a short time exposure to the stratum comeum of the epidermis, the cytotoxic effects of the test item on EpiDerm™, a reconstituted three-dimensional human epidermis model, were determined. Hereby, the test item was applied topically.

Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT after a 3 min and 60 min exposure period and compared to those of the concurrent negative controls. The tissues were treated with each dose group in duplicate. The positive control was 8 N Potassium Hydroxide and the negative control was distilled water.

In this study under the given conditions the test item showed no corrosive effects. The relative mean tissue viability after 3 min treatment was >= 50% and after 60 min. treatment >= 15%. The test item is therefore classified as "non corrosive".

Reference 2

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2007-11-12 to 2007-11-20

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

adopted 24th April, 2022

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Version / remarks:

2004

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2500 (Acute Dermal Irritation)

Version / remarks:

August 1998

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Remarks:

New Zealand White Hsd If:NZW

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Winkelmann GmbH, D-33178 Borchen
- Age at study initiation: not specified
- Weight at study initiation: 2.1 - 3.4 kg
- Fasting period before study: overnight, after administration of the test item the food was withheld for for further 3- 4 h
- Housing: ABS-plastic rabbit cages, floor 4200 cm2
- Diet (ad libitum): Altromin 2123 maintanenance diet for rabbits, specific pathogen-free
- Water (ad libitum): tap water
- Acclimation period: at least 5 days
- Microbiological status: SPF

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 +/- 3
- Humidity (%): 55 +/- 10
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Remarks:

24 hours before the test, fur was removed from the dorsal area of the trunk by clipping.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 mL

Duration of treatment / exposure:

4 h

Observation period:

72 h after patch removal
120 h after patch removal if symptoms present after 72 h

Number of animals:

3

Details on study design:

TEST SITE
- Area of exposure: 6 cm2
- Type of wrap: gauze patch, held in place with non-irritating tape and fixed with semi-occlusive dressing.

REMOVAL OF TEST SUBSTANCE: not specified. It was stated that the test item was stuck to the application site

OBSERVATION TIME POINTS: 1, 24, 38 nd 72 h

SCORING SYSTEM: according to Draize (1944)

1. Erythema and Eschar Formation

No erythema 0
Very slight erythema (barely perceptible) ..................................................................................1
Well defined erythema ................................................................................................................ 2
Moderate to severe erythema .................................................................................................... 3
Severe erythema (beef redness) to eschar formation preventing grading of erythema ........ 4
Maximum possible: 4

2. Oedema Formation

No oedema 0
Very slight oedema (barely perceptible) .....................................................................................1
Slight oedema (edges of area well defined by definite raising) ............................................... 2
Moderate oedema (raised approximately 1 mm) .......................................................................3
Severe oedema (raised more than 1 mm and extending beyond area of exposure) ...............4
Maximum possible: 4

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.67

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

probability of mild irritation

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 120 h

Remarks on result:

probability of moderate irritation

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.67

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

probability of mild irritation

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Reversibility of any observed effect: changes fully reversible within 5 days

Other effects:

The test item showed slightly irritant, but no corrosive effects on the intact skin of 3 rabbits after a contact time of 4 h.

The scores for animals 1, 2 and 3 were (0.67, 1, 0.67) for erythema and (0, 0, 0) for edema.

Animals No. 1 and 3: All symptoms were fully reversible within 72 hours after removing the patch.

Animal No. 2: All syptoms were fully reversible within 120 hours after removing the patch.

There were no significant body weight changes during the contact and observation period.

Interpretation of results:

GHS criteria not met

Conclusions:

Considering the reported data of this irritation study, it can be stated that the test item showed slightly irritant effects, which were fully reversible.

Executive summary:

The skin irritation potential of the test item was assayed in an in vivo skin irritation study according to OECD 404 in three New Zealand rabbits. 0.5 mL of the test item was applied per test side (dorsal area of the trunk), the untreated side served as control. The exposure period was 4 h. A semiocclusive dressing was used. The test sides were observed 1, 24, 48 and 72 h after patch removal for erythema and oedema.

The test item showed slightly irritant, but no corrosive effects on the intact skin of 3 rabbits after a contact time of 4 h. The scores for erythema and edema were (0.67, 0) for animal 1, (1, 0) for animal 2 and (0.67, 0) for animal 3.

Animals No. 1 and 3: All symptoms were fully reversible within 72 hours after removing the patch.

Animal No. 2: All symptoms were fully reversible within 120 hours after removing the patch.

There were no significant body weight changes during the contact and observation period.

The test item was concluded to be slightly irritant, with full reversibility.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2007-11-20 to 2007-11-25

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Version / remarks:

August 1998

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Version / remarks:

2004

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

April 2002

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Remarks:

New Zealand White Hsd If:NZW

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Harlan Winkelmann GmbH, D-33178 Borchen
- Age at study initiation: not specified
- Weight at study initiation: 2.6 - 2.7 kg
- Housing: ABS-plastic rabbit cages, floor 4200 cm2
- Diet (ad libitum): Altromin 2123 maintanenance diet for rabbits, specific pathogen-free
- Water (ad libitum): tap water
- Acclimation period: at least 5 days
- Microbiological status: SPF

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 +/- 3
- Humidity (%): 55 +/- 10
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1 mL

Duration of treatment / exposure:

not applicable, no washing;
lids were gently hold together for 1 sec

Observation period (in vivo):

72 h

Number of animals or in vitro replicates:

3 females

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: no

SCORING SYSTEM: according to Draize (1944)

Endpoint Description Range
Cornea degree opacity and ulcerations 0–4
Iris swelling, hyperaemia 0–2
Conjunctivae redness, vessel discernibility 0–3
Chemosis swelling, lids closed/open 0–4

TOOL USED TO ASSESS SCORE: fluorescein

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.67

Max. score:

3

Reversibility:

fully reversible within: 72 h

Remarks on result:

probability of mild irritation

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 48 h

Remarks on result:

probability of weak irritation

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 48 h

Remarks on result:

probability of weak irritation

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

probability of weak irritation

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Animal No. 1 showed redness (grade 1), discharge (grade 2) as well as chemosis of the conjuctivae (grade 1) 1 h after instillation of the test item. 24 h post-treatment redness and chemosis (grade 1) were observed. After 48 h, redness (grade 1) was observed. After 72 h, no symptoms were obseved. Thus, the changes were fully reversible within 72 hours post instillation.

Animals No. 2 and 3 showed redness (grade 1) as well as chemosis of the conjuctivae (grade 1) 1 h after instillation of the test item. 24 h post-treatment redness (grade 1) was observed. After 48 h, no symptoms were obseved. Thus, the changes were fully reversible within 48 hours post instillation.

The scores were (conjuctivae, chemosis, cornea opacitiy, iris):
Animal 1: (0.67; 0.33; 0; 0)
Animal 2: (0.33; 0 ;0; 0)
Animal 3: (0.33; 0 ;0; 0)

Other effects:

There were no significant body weight changes during the observation period.

Interpretation of results:

GHS criteria not met

Conclusions:

Considering the reported data of this irritation study, it can be stated that the test item produced slightly irritating, but no corrosive effects in the eyes of rabbits. All signs of irritation were fully reversible.

Executive summary:

The skin irritation potential of the test item was assayed in an in vivo eye irritation study according to OECD 405 in three female New Zealand White rabbits (HsdIf:NZW). 0.1 mL of the test item was instilled in the lower conjuctival sac of one eye of each animal, the untreated contralateral eye served as control. The animals were observed for 72 h for ocular lesions in the cornea, iris and conjuctivae.

Observations were scored and compared to the control eyes 1, 24, 48 and 72 h post-dose. The test item showed slightly irritant, but no corrosive effects on the eyesof 3 rabbits.

Animal No. 1 showed redness (grade 1), discharge (grade 2) as well as chemosis of the conjuctivae (grade 1) 1 h after instillation of the test item. 24 h post-treatment redness and chemosis (grade 1) were observed. After 48 h, redness (grade 1) was observed. After 72 h, no symptoms were obseved. Thus, the changes were fully reversible within 72 hours post instillation.

Animals No. 2 and 3 showed redness (grade 1) as well as chemosis of the conjuctivae (grade 1) 1 h after instillation of the test item. 24 h post-treatment redness (grade 1) was observed. After 48 h, no symptoms were obseved. Thus, the changes were fully reversible within 48 hours post instillation.

The scores were (conjuctivae, chemosis, cornea opacitiy, iris): (0.67; 0.33; 0; 0) for animal 1, (0.33; 0 ;0; 0) for animal 2 and (0.33; 0 ;0; 0) for animal 3.

There were no significant body weight changes during the observation period. Further, no corneal lesions were found in any animal at the final reading.

The test item was concluded to be slightly irritant to the eye, with full reversibility.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation/ corrosion

In vivo, according to OECD 402

The skin irritation potential of the test item was assayed in an in vivo skin irritation study according to OECD 404 in three New Zealand rabbits. 0.5 mL of the test item was applied per test side (dorsal area of the trunk), the untreated side served as control. The exposure period was 4 h. A semiocclusive dressing was used. The test sides were observed 1, 24, 48 and 72 h after patch removal for erythema and oedema.

The test item showed slightly irritant, but no corrosive effects on the intact skin of 3 rabbits after a contact time of 4 h. The scores for erythema and edema were (0.67, 0) for animal 1, (1, 0) for animal 2 and (0.67, 0) for animal 3.

Animals No. 1 and 3: All symptoms were fully reversible within 72 hours after removing the patch.

Animal No. 2: All symptoms were fully reversible within 120 hours after removing the patch.

There were no significant body weight changes during the contact and observation period.

The test item was concluded to be slightly irritant, with full reversibility and not requiring classification.

 

In vitro, according to OECD 431

The skin corrosivity potential of the test item was analysed according to OECD 431. Since corrosive chemicals are cytotoxic after a short time exposure to the stratum comeum of the epidermis, the cytotoxic effects of the test item on EpiDerm™, a reconstituted three-dimensional human epidermis model, were determined. Hereby, the test item was applied topically. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT after a 3 min and 60 min exposure period and compared to those of the concurrent negative controls. The tissues were treated with each dose group in duplicate. The positive control was 8 N Potassium Hydroxide and the negative control was distilled water.

In this study under the given conditions the test item showed no corrosive effects. The relative mean tissue viability after 3 min treatment was >= 50% and after 60 min. treatment >= 15%. The test item is therefore classified as "non corrosive".

 

Eye irritation

in vivo, according to OECD 405

The eye irritation potential of the test item was assayed in an in vivo eye irritation study according to OECD 405 in three female New Zealand White rabbits (HsdIf:NZW). 0.1 mL of the test item was instilled in the lower conjuctival sac of one eye of each animal, the untreated contralateral eye served as control. The animals were observed for 72 h for ocular lesions in the cornea, iris and conjuctivae.

Observations were scored and compared to the control eyes 1, 24, 48 and 72 h post-dose. The test item showed slightly irritant, but no corrosive effects on the eyes of 3 rabbits.

The scores were (conjuctivae, chemosis, cornea opacitiy, iris): (0.67; 0.33; 0; 0) for animal 1, (0.33; 0 ;0; 0) for animal 2 and (0.33; 0 ;0; 0) for animal 3. The changes were fully reversible with 72 h for animal 1 and 48 h for animals 2 and 3. No corneal lesions were found in any animal at the final reading.

There were no significant body weight changes during the observation period. 

The test item was concluded to be slightly irritant to the eye, with full reversibility and not requiring classification.

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008.

 

Skin: Based on available data on skin irritation, the test item does not require classification according to Regulation (EC) No 1272/2008 (CLP), as amended for the eighteenth time in Regulation (EU) 2022/692.

 

Eye: Based on available data on eye irritation, the test item does not require classification according to Regulation (EC) No 1272/2008 (CLP), as amended for the eighteenth time in Regulation (EU) 2022/692.